Figure 5.

TRMT112 single amino acid mutant interactions with MTases confirm similarities, but also reveal differences in the way TRMT112 interacts with its partners. U2OS cells were transfected with plasmids encoding for EGFP-tagged N6AMT1 (A), WBSCR22 (B), METTL5 (C), TRMT11 (D), ALKBH8 (E), THUMPD3 (F), THUMPD2 (G) and EGFP (H), along with plasmids encoding for wt or mutated E2Tag-containing TRMT112. Immunoprecipitation was performed with GFP-Trap beads 48 h later using EGFP-tagged MTase as a bait, and samples were subsequently analysed using Western blot. Upper three images of A-H panel represent input of total soluble cell lysate, whereas lower three images represent immunoprecipitated material. Both sections were analysed using antibodies against EGFP, E2Tag and TRMT112. Images of results with anti-TRMT112 depict both recombinant (upper band) and endogenous (lower band marked with asterisk *) protein.